TY  - JOUR
AU  - Meurs, Romane
AU  - De Matos, Mara
AU  - Bothe, Adrian
AU  - Guex, Nicolas
AU  - Weber, Tobias
AU  - Teleman, Aurelio
AU  - Ban, Nenad
AU  - Gatfield, David
TI  - MCTS2 and distinct eIF2D roles in uORF-dependent translation regulation revealed by in vitro re-initiation assays.
JO  - The EMBO journal
VL  - 44
IS  - 3
SN  - 0261-4189
CY  - Hoboken, NJ [u.a.]
PB  - Wiley
M1  - DKFZ-2025-00043
SP  - 854-876
PY  - 2025
N1  - 2025 Feb;44(3):854-876
AB  - Ribosomes scanning from the mRNA 5' cap to the start codon may initiate at upstream open reading frames (uORFs), decreasing protein biosynthesis. Termination at a uORF can lead to re-initiation, where 40S subunits resume scanning and initiate another translation event downstream. The noncanonical translation factors MCTS1-DENR participate in re-initiation at specific uORFs, but knowledge of other trans-acting factors or uORF features influencing re-initiation is limited. Here, we establish a cell-free re-initiation assay using HeLa lysates to address this question. Comparing in vivo and in vitro re-initiation on uORF-containing reporters, we validate MCTS1-DENR-dependent re-initiation in vitro. Using this system and ribosome profiling in cells, we found that knockdown of the MCTS1-DENR homolog eIF2D causes widespread gene deregulation unrelated to uORF translation, and thus distinct to MCTS1-DENR-dependent re-initiation regulation. Additionally, we identified MCTS2, encoded by an Mcts1 retrogene, as a DENR partner promoting re-initiation in vitro, providing a plausible explanation for clinical differences associated with DENR vs. MCTS1 mutations in humans.
KW  - DENR-MCTS1 (Other)
KW  - In Vitro Translation (Other)
KW  - Re-Initiation (Other)
KW  - eIF2D (Other)
KW  - uORF (Other)
LB  - PUB:(DE-HGF)16
C6  - pmid:39748120
DO  - DOI:10.1038/s44318-024-00347-3
UR  - https://inrepo02.dkfz.de/record/296113
ER  -